Background: Necrotizing enterocolitis (NEC) is the leading gastrointestinal cause of death of premature neonates. We have previously shown that this hyperinflammatory state persists even post-recovery. We hypothesize that recovered patients with NEC will have a decreased hyperinflammatory response when the anti-inflammatory medication mesalamine (5-ASA) is administered even when exposed to in vitro NEC induction. Methods: Enteroids were generated and subjected to in vitro NEC induction. One half were subjected to 5-ASA treatment. Tumor necrosis factor-alpha (TNF-α) and interleukin 8 (IL-8) were evaluated via RT-qPCR. Mice underwent in vivo NEC induction, one group was given 5-ASA 50 mg/kg 12 h before the start of NEC induction. The intestine was harvested and assessed for hyperinflammatory markers and histological grading was performed. Results: Recovered NEC enteroids treated with 5-ASA during NEC induction show a significant decrease in inflammatory markers compared with control (p = 0.0014 TNF-α, downtrend IL-8). Active NEC enteroids treated with 5-ASA during in vitro NEC induction show a significant decrease in TNF-α RT-qPCR (p = 0.0443) and IL-8 RT-qPCR (p = 0.0265). In mice that received 5-ASA 50 mg/kg before in vivo NEC induction, there is a significant decrease in both TNF-α (p = 0.0114) and IL-8 (p = 0.0051). Conclusion: Enteroids and mice exposed to 5-ASA have a significant decrease in inflammatory markers. This decrease despite NEC induction in both enteroids and mice may demonstrate the impact that anti-inflammatory agents could have on treatment for NEC. This could be important given the robust hyperinflammatory response to a second hit after recovery and may impact the trajectory of an illness post-recovery from NEC.

Keywords: 5-ASA; enteroids; hyperinflammatory; necrotizing enterocolitis; recovery.

背景： 溃疡性坏死性小肠结肠炎（NEC）是早产儿胃肠死亡的主要原因。我们之前的研究显示，这种过度炎症状态即使在恢复后仍然存在。我们假设，在体外NEC诱导过程中给予抗炎药物美沙拉秦（5-ASA）治疗时，已经从NEC中康复的患者将表现出较低的过度炎症反应。 方法： 生成肠类器官并进行体外NEC诱导。一半接受5-ASA治疗。通过RT-qPCR评估肿瘤坏死因子α (TNF-α) 和白细胞介素8 (IL-8)。小鼠体内接受NEC诱导，一组在开始NEC诱导前12小时给予50 mg/kg的5-ASA。收集肠道并检测过度炎症标记物，并进行组织学分级。 结果： 在体外NEC诱导过程中用5-ASA治疗的康复NEC肠类器官显示出TNF-α和IL-8炎症标志物显著降低（p = 0.0014 TNF-α，IL-8趋势下降）。在体内NEC诱导前接受5-ASA的小鼠中，在体外NEC诱导过程中用5-ASA治疗的活性NEC肠类器官显示TNF-α RT-qPCR (p = 0.0443) 和 IL-8 RT-qPCR (p = 0.0265) 显著降低。在接受5-ASA 50 mg/kg后体内NEC诱导的小鼠中，两种 TNF-α (p = 0.0114) 和 IL-8 (p = 0.0051) 均显著下降。 结论： 接受5-ASA治疗的肠类器官和小鼠在炎症标志物方面表现出显著降低。即使是在体外NEC诱导过程中，这种减少也可能显示抗炎药物对NEC治疗的影响。考虑到恢复后对第二次打击产生的强烈过度炎症反应，这可能会影响从NEC康复后的疾病轨迹。

关键词：5-ASA；肠类器官；过度炎症；坏死性小肠结肠炎；康复。